Process Biochemistry 47 (2012) 17611772

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Process Biochemistry
journal homepage: www.elsevier.com/locate/procbio

Review

Probiotic meat products and human nutrition

a, Danuta Koozyn-Krajewska , Zbigniew J. Dolatowski b
a b

Warsaw University of Life Sciences-SGGW, Nowoursynowska 159c, 02-776 Warszawa, Poland University of Life Sciences in Lublin, Skromna 8, 20-704 Lublin, Poland

a r t i c l e

i n f o

a b s t r a c t
Raw cured and ripened meat products have been traditionally manufactured using the fermentation of native or added carbohydrates by lactic acid bacteria found in meat or in its environment. The commercial application of probiotic microorganisms in dry fermented meat products is not yet common. Probiotic bacterial strains that can be used in the manufacturing of dry fermented meat products should be capable of surviving in conditions found in fermented products; furthermore, they should dominate other microorganisms found in the nished product. The initial number of microorganisms in sausage lling or on the surface of ham or loin cannot be reduced as in milk pasteurization, for example. Therefore, the choice of appropriate microorganisms is important. Probiotic meat products are a relatively new and not very well recognized eld of meat industry, but the most important issue is to nd a compromise between technological aspects, safety, quality and health-benecial effects of food. Therefore, the object of this review is on the one hand to analyze technological possibilities and quality parameters of probiotic meat products, and on the other hand to discuss risks and benets of probiotic meat used in human nutrition. 2012 Elsevier Ltd. All rights reserved.

Article history: Received 8 May 2012 Received in revised form 14 September 2012 Accepted 23 September 2012 Available online 8 October 2012 Keywords: Probiotics Meat, Technology Quality Benets Risk

Contents 1. 2. 3. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Probiotic microorganisms and health benets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Fermented meats in human nutrition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.1. Microorganisms as starter cultures for processed meat products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.2. Types of fermented meat products (hams, loins, sausages) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Meat as a potential carrier of probiotic bacteria technological aspects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Microbiological, sensory and physicochemical parameters of probiotic meat products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.1. Survival of probiotic bacteria used in the production of dry fermented meat products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.2. Lipid and protein oxidation in probiotic products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.3. Sensory properties of probiotic fermented meat products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Probiotic meat products in human nutrition benets and risks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.1. Human studies using probiotic fermented meats . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2. Safety of probiotic meat products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.3. Biogenic amines in probiotic meat products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusions and future trends . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1761 1762 1762 1762 1763 1764 1766 1766 1766 1767 1768 1768 1768 1768 1769 1770

4. 5.

6.

7.

1. Introduction The trend today is the development of novel food for special health use, called functional food, to promote human health and well-being of consumers [5,96]. Meat and meat products are one

Corresponding author. Tel.: +48 22 5937069; fax: +48 22 5937069. E-mail addresses: danuta kolozyn krajewska@sggw.pl (D. Koozyn-Krajewska), zbigniew.dolatowski@up.lublin.pl (Z.J. Dolatowski). 1359-5113/$ see front matter 2012 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.procbio.2012.09.017

of the most important components of human nutrition. Raw cured and ripened meat products (different kinds of sausage and smoked cured meat products) have been traditionally produced using the fermentation process, a guided decomposition process involving meat intrinsic enzymes and enzymes of microbiological origin. The traditional production of raw meat products is based on the fermentation of native or added carbohydrates by lactic acid bacteria (LAB) found in meat or in its environment. Numerous substances form during fermentation, such as lactic acid, acetic acid, alcohols, aldehydes, ketones and bacteriocins, which affect the quality, safety and

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storage stability of a meat product. During the last 40 years it has been quite a common practice in manufacturing raw ripened meat products to apply starter cultures whose composition is extremely varied [41,42,53,55,102]. Probiotic products are the fastest growing area of novel food product development. Probiotic bacteria strains are successfully used in the production of processed milk products, cereal products [52,82], bread-making [74] and vegetable and fruit juices [58]; however, they have not been used in raw ripening meat products. The commercial application of probiotic microorganisms in dry fermented meat products is not yet common. The main technological problem is meat matrices: on the one hand they are adequate for the growth and survival of probiotic bacteria as they are usually not heated, but on the other hand, bacteria may be inactivated due to the high content of curing salt, low pH and low water activity. A serious issue is also the native microorganisms present in raw meat [53,55,57,86]. In the production of probiotic meat products, bacterial strains with documented probiotic properties only should be used. As these are usually strains isolated from human intestines, they should be able to compete with the bacteria found in meat, an environment not natural for them. They are also supposed to be capable of surviving the process of fermentation and drying and that of cooling and storage and also of multiplying to the number considered to bring health-benecial effects [8,80]. When producing meat products, the initial number of microorganisms in sausage lling or on the surface of ham or loin cannot be reduced as in milk pasteurization, for example. Therefore, the choice of appropriate microbial strains will be important. One obvious technological possibility is to use bacteria closely related to the meat environment that have suitable properties and health-benecial characteristics. Bacteria with probiotic properties can be obtained as natural isolates from fermented products (e.g. dry sausages or hams) or from existing commercial meat starter cultures. Another possibility is the application of strains with documented probiotic properties as starter cultures [20,53,78,80,8486]. Probiotic meat products are a relatively new and not very well recognized eld of meat industry, but the most important issue is to nd a compromise between technological aspects, safety, quality and health-benecial effects of food. Therefore, the object of this review was on the one hand to analyze technological possibilities and quality parameters of probiotic meat products, and on the other hand to discuss risks and benets of probiotic meat use in human nutrition.

the large intestine and confer a health benet on the host [81]. Throughout the years, several selection criteria for probiotics have been suggested [5,20,73,79]: human origin, survival of the passage through the upper gastrointestinal tract (resistance to gastric acid, bile salts, and proteolytic enzymes, activity during transit from the stomach to the colon), interactions with the resident microbiota (ability to adhere to the intestinal epithelium and colonize the colon, no signicant effect on the dominant microbiota), benecial effect on the host (probiotichost interactions, prevention of diseases), antagonism against pathogenic bacteria by competitive exclusion, resistance to technological processing (e.g. effects of the food matrix, food ingredients and mechanical and heat treatment) and storage, safety in human use and excellent tolerance. It should, however, be noted that probiotic strain characteristics, such as survival of the passage through the GI tract, are not sufcient to consider a microbial strain probiotic. According to the WHO/FAO denition, the main criterion for a probiotic strain should be the fact that it confers a health benet on the host, which can only be demonstrated through well-designed, randomized, double-blind, placebo-controlled, multi-center human trials whose the results are published in peer-reviewed international scientic journals [79]. Probiotics may be available in different forms, such as foods, mainly in the fermented state, and pharmaceutical products, mainly as capsules or in microencapsulated forms. The rst step in every process of probiotic food product development is the identication and characterization of the probiotic strain at the genus, species, and strain level, and next steps are investigations of physiological characteristics and technological requirements, mechanisms of action and demonstration of human benets [19,20,39]. Probiotic foods comprise between 60 and 70% of the total functional food market. A continued growth is observed mainly among dairy-type probiotic foods, but also in the range of non-dairy probiotic food products, such as fermented meats and vegetable and fruit juices. Taking into account the wide range of potential (fermentable) substrates and the different conditions in which LAB strains may reveal functional performance, it can be expected that developments toward new food-based probiotics will proceed further in the future [39,96].

3. Fermented meats in human nutrition 3.1. Microorganisms as starter cultures for processed meat products

2. Probiotic microorganisms and health benets The expression probiotic was probably rst dened by Kollath in 1953 [39], when he suggested the term to denote all organic and inorganic food complexes as probiotics, in contrast to harmful antibiotics, to use such food complexes as supplements. Lilly and Stillwell in 1965 [87] proposed probiotics to be microorganisms promoting the growth of other microorganisms. To act as a safe probiotic microorganism a strain should be of species and genera normally present in the human gastrointestinal tract. Probiotics are lactic acid bacteria or bidobacteria, mainly Gram-positive Lactobacillus species. The genus Bidobacterium shares some phenotypic features with the typical LAB, but traditionally and also for practical purposes it is still considered to belong to LAB. Also enterococci have been proposed as probiotics; however, considering their potential pathogenicity and antibiotic resistance gene transfer, they are not recommended for human use [39,73,79]. In general, health benets of probiotic foods are based on the presence of selected strains of lactic acid bacteria that, having passed through the stomach and the small intestine, survive in The preservation of meat by fermentation through native microorganisms has been used for thousands of years. During those transformations numerous substances form in the products, such as lactic acid, pyruvic acid, alcohols, aldehydes, ketones, and carboxylic acids that shape the quality and storage stability of a product [41,43,60]. Nowadays, it is quite a common practice in manufacturing raw cured ripened products to apply starter cultures to ensure a standard quality of the fermentation process. Meat starter cultures are preparations which contain live or dormant forms of microorganisms that develop desired metabolic activity in the fermentation substrate. As a rule, but not necessarily, the organisms multiply in this substrate [37]. Starter cultures are facultative heterofermentative strains that produce lactic acid from hexoses (glucose). They also produce both lactic acid and acetic acid from pentoses (arabinose, xylose) [22]. The selection of a suitable strain for each product is essential (Table 1). The most common LAB strains in starter cultures are Lactobacillus sakei, Lactobacillus curvatus, Lactobacillus plantarum,

 D. Koozyn-Krajewska, Z.J. Dolatowski / Process Biochemistry 47 (2012) 17611772 Table 1 Microorganisms as starter cultures for processed meat products [partly based on 55]. Microorganisms in starter cultures Lactic acid bacteria Lactobacillus plantarum Lactobacillus sake Lactobacillus pentosus Lactobacillus casei Lactobacillus curvatus Lactobacillus alimentarius Pediococcus acidilacti Pediococcus pentosaceus Gram-positive cocci Staphylococcus carnosus Staphylococcus xylosus Micrococcus varians Type of action and technological changes in meat Lactic acid production, Inhibition of growing of undesired bacteria, Acceleration of food colors reaction (completion of the curing process), Drying process acceleration, Flavor and aroma development

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Nitrates and nitrites reduction, Using up the oxygen, Decomposition of peroxides, Lipolysis, Color stabilization, Delaying rancidity, Flavor and aroma development Using up the oxygen, Decomposition of peroxides, Color stabilization, Delaying rancidity, Flavor and aroma development Using up the oxygen, Decomposition of peroxides, Proteolysis, Lipolysis Flavor and aroma development

Yeasts Debaryomyces hansenii Candida famata

Moulds Penicillium nalgiovense Penicillium camamberti Penicillium chrysogenum

Lactobacillus lactis, Lactobacillus casei, Pediococcus pentosaceus, and Pediococcus acidilactici [3,41,104]. L. sakei, L. curvatus, L. plantarum, the most frequently isolated LAB from fermented sausages adapt themselves to the environment that prevails in the sausage during fermentation; they control the ripening process and the growth of undesired microbiota. Pediococci are less frequently isolated from European fermented sausages, but they are more common in fermented sausages from the United States where they are deliberately added as starter cultures to accelerate the acidication of meat batter [55]. Different LAB strains produce intrinsically different amounts of lactic acid, which needs to be considered while choosing an LAB strain for a specic meat product; however, several other factors also affect the rate and amount of acid formation. The bacterial growth rate is higher at higher temperatures and water activity. Lactic acid production depends on sausage diameter (lower or higher oxygen level) [22,41]. 3.2. Types of fermented meat products (hams, loins, sausages) A raw cured sausage is a mixture of different meat grades preserved by curing, stuffed into casings and microbiologically fermented with salt, nitrate or nitrite, sugar and spices (mostly garlic and black pepper) added [43]. For the fermentation process to be suitable and to obtain a desired lactic acid quantity, the sausage has to be stored at a temperature appropriate for lactic acid bacteria (2737.8 C) for 1015 h so that the bacteria begin to grow and transform carbohydrates into lactic acid [20]. Manufacturing raw cured sausages has a very long tradition and its origin is to be sought in the Roman times in the Mediterranean area. In terms of quantity and quality, the most important European countries which manufacture raw cured sausages are Germany, Italy, Spain, France, the Netherlands, Belgium and Austria. In Germany, most sausages are smoked, and in Mediterranean countries, in France and Hungary, raw cured air-dried sausages are principally manufactured, with signicant mould development on their surface. The addition of starter culture bacteria is necessary

for the production of raw cured sausages. They initiate the process of fermentation, control it and ensure microbiological safety of the nished product [6,8,20,41,54,75,78,80,104]. In the past, raw cured sausages were manufactured only in the cool and cold time of the year and, therefore, it was possible to avoid the risk of spoilage. The development of ripening rooms made it possible to manufacture raw cured sausages of the same quality throughout the year, regardless of ambient conditions. Currently, one of the most important parameters in the production of raw cured sausages is to shorten ripening time and obtain the dened sensory characteristics during that time. The trend to manufacture faster ripening products (pH within a range of 5.25.3 or slightly lower after 24 h), ready for sale within approx. 14 days and sometimes even earlier results from economic reasons, so there is a demand for fast-acidifying starter cultures. Such products can be acidied by L. curvatus or L. sakei bacteria [36,55]. Compared with typical, especially long-ripening sausages, fast ripening raw cured sausages are characterized by a less expressed aroma and a slightly paler color. This is caused by the fact that as a result of a fast decrease in the pH value, Staphylococci cause transformations that contribute to the achievement of a characteristic aroma and color. Individual Staphylococcus strains differ from one another e.g. by producing proteases (protein decomposing enzymes), peptidases (peptide decomposing enzymes) and lipases (fat decomposing enzymes) with different activities [41,43,64,78,109]. Salami-type raw cured sausages are a wide group of products which differ in raw material composition, ripening time, external appearance and avor. The original recipe for the salami-type raw cured sausage, initially made from donkey meat, was developed in Italy. It is widespread in such countries as Hungary, Germany, Italy, France or Spain. Raw cured fermented sausages are also manufactured in Poland, but in relatively small quantities [9,55,78,109]. The avor of fermented sausages is inuenced by several factors: source, quantity and type of ingredients (e.g. meat, salt and spices), temperature, processing time, smoking and choice of the starter culture. Basic avor results from the interaction of taste (mainly determined by lactic acid production, peptides and free amino acids) and aroma [60,61,91]. For cured meat products, different spices and additives are also used. Ground meat is preserved due to the fermentation process and air-drying. The low pH caused by fermentation and decreased water activity through drying with relatively high salt concentration creates an unfavorable environment for pathogenic bacteria growth. Such cured meat products are nished products ready for consumption and they do not require heat treatment [55]. Ripened products made from raw ham were manufactured as early as in the Middle Ages and their numerous sorts were known in Central and Southern Europe. Also in China Jinhua ham is produced and is one of the most famous traditional meat products [114]. The ham is principally a cut from a hind leg. Raw ripened hams are a group of meat products more difcult to manufacture than others. In such products, the selection of the raw material plays an important role and diffusion of salt components is more difcult, which causes undesirable microbial growth leading to product decay or health hazard. Furthermore, the different consistency of individual muscles creates technological problems. During curing, fermentation, ripening and storage, sarcoplasmic and myobrillar meat proteins are hydrolyzed by microbial proteases and endogenous meat enzymes. Polypeptides, peptides and free amino acids are among the compounds formed through proteolysis [91]. Different fat quantities on the surface make skin-drying uneven and lead to aw differences between muscles and thereby an excessive microbial growth or suppression of growth. Hams are eliminated from the market as a result of decay symptoms, such as soft consistency, chromatosis, and even perforation caused by excessive gas

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production. Ripened ham manufacturing requires a careful choice of raw materials and control of the ripening process [55,109,114].

4. Meat as a potential carrier of probiotic bacteria technological aspects Dry fermented meat products are usually not heated or only mildly heated, which is adequate for the transfer of probiotic bacteria into the human gastrointestinal tract [3]. In addition, it is believed that the meat matrix supports the survival of probiotic lactobacilli through the gastrointestinal tract. The probiotic culture should be well-adapted to the conditions in fermented products (ham, loin, sausages) and become dominant in the nal products which contain the high natural background microbiota. However, the potential negative impact of the meat environment on cell viability must be taken into account, in particular with respect to the high content of curing salt and its low pH and water activity due to acidication and drying. The choice of appropriate microorganisms is very important due to cell viability in a fermented meat environment which is most likely strain-dependent [20,103]. Manufacturing probiotic meat products is much more difcult than that of other probiotic products and such products are at their initial development stage. The reasons are to be found in raw material characteristics. Probiotic bacteria strains that can be used in the manufacturing of fermented meat products should survive in fermented products and they should dominate other microorganisms found in the product. One possibility is the use of bacterial strains obtained by screening natural sausage isolates or meat starter cultures for probiotic properties (Table 2). Since the microora of spontaneously ripened European-style raw fermented sausages is usually dominated by more competitive strains of the L. sakei/curvatus group, the suitability of new probiotic cultures must be veried for each individual product type [57]. Most LAB strains have a high degree of hydrophobicity, indicating that these microorganisms have a probiotic potential. Some bacterial strains isolated from Iberian dry fermented sausages have been characterized as probiotic and these strains were considered safe for use. However, the microorganisms used as probiotic cultures should demonstrate good adaptation to the real conditions present in the fermented sausage and become dominant in the nal product, since it contains large natural background microbiota. Therefore, the growth of LAB in Iberian sausages depends on their ability to tolerate salt concentration (NaCl, 2.53.5%), pH values (4.75.6) and water activity (0.840.85) in the products during the ripening process [85,86]. The most frequently isolated LAB from dry sausages processed using different technologies are L. sakei, L. curvatus, L. plantarum, Lactobacillus fermentum, Lactobacillus brevis, P. acidilactici and Pediococcus pentoseaceus [35,37,90]. In Iberian dry fermented sausages, P. pentosaceus, P. acidilactici, Lactococcus lactis and L. brevis have been isolated as the predominant LAB, together with sporadic isolates of Leuconostoc mesenteroides, L. plantarum and L. curvatus [8]. The importance of using dominant strains is related to the demand for higher numbers of viable cells at the time of consumption, which is a prerequisite for the culture to ensure benecial effects on the host. In this aspect, Klingberg et al. [54] reported the selection of L. plantarum and L. pentosus naturally present in a Scandinavian-type fermented sausage as promising candidates for probiotic meat starter cultures. Papamanoli et al. [75] found that L. plantarum strains and L. curvatus strains isolates from Greek dryfermented sausages were resistant to 0.3% bile salts. Arihara [5] reported that probiotic strains (Lactobacillus acidophilus, L. casei, L. paracasei, Bidobacterium spp.) are suitable for meat fermentation, and Sameshima et al. [88] demonstrated the potential usefulness of Lactobacillus rhamnosus and L. paracasei subsp. Paracasei strains

in meat fermentation. Erkkila et al. [25] and Erkkila and Petaja [23] used L. rhamnosus for dry sausage manufacturing. Andersen [4] fermented dry sausages using a mixture of one commercial starter culture and L. casei or a mixture of the same starter and probiotic B. lactis. Pidcock et al. [78] reported that non-traditional meat starter cultures (probiotic strains) can be used to maintain safety and sensory properties of Hungarian salami. Lactobacilli are of the highest importance in meat fermentation because of their ability to provide rapid and effective fermentation and acidication, thus preserving the sausages from the development of spoilage and pathogenic bacteria. Therefore, they are often used as starters in dry fermented sausage production. Combining probiotic potential and technological performance of Lactobacillus strains would lead to interesting probiotic starters for use in novel dry fermented sausages [77]. Alternatively, the performance of strains with documented health-promoting properties may be investigated in a fermented meat environment. Since such strains are usually human intestinal isolates, they should be able to compete with the natural meat microbiota in an environment which is not their natural habitat, to survive the fermentation and drying process as well as refrigeration and storage conditions and, preferably, to grow to numbers that enable the occurrence of health-promoting effects [20]. Many scientists have recently proposed the use of probiotic bacteria of various origin in dry fermented products (Table 2). Meat curing is adding nitrate and/or nitrite with salt to meat. Sodium chloride is one of the oldest and most common agents for food preservation. The salt concentration used in fermented products inhibits the growth of many undesirable microorganisms and thereby promotes the growth of more salt tolerant lactic acid bacteria. NaCl also has a profound inuence on the water binding properties of meat by inuencing the solubilization and diffusion of myobrillar proteins. Pathogenic and spoilage microorganisms are generally inhibited in fermented products due to a combination of several factors: high salt, presence of nitrite curing salt, low aw , reduced redox potential due to the removal of residual O2 by Gram-negative bacteria in the meat, growth of competitive starter cultures and pH decrease [40,42,113]. However, nitrate becomes active only after being microbially reduced to nitrite. The basic function of nitrite is the production of the characteristic pink color Nitrite contributes to the development of the characteristic color and avor of cured meat products, provides inhibition of spoilage and pathogenic bacteria, such as Clostridium botulinum, and controls rancidity and off-avor development in the products [17,34]. The biochemical basis of red color in meats is well established, and depends on the concentration and redox state of hem pigments in meat [7,29,66]. Myoglobin exists in a muscle in three forms: native myoglobin (MbFe2+ ), oxymyoglobin (MbO2 ) and metmyoglobin (MbFe3+ ). The pigment responsible for the bright red color of cured meat is a ferrous complex of myoglobin (MbFe2+ ) and a ligand, nitric oxide (NO), called nitrosylmyoglobin (MbFe2+ NO). The formation of nitrosylmyoglobin requires two important factors: low pH and a reducing agent. However, nitrosylmyoglobin can be oxidized to brown metmyoglobin (MbFe3+ ) by oxygen and certain other oxidizing agents, such as nitrite. The addition of reducing agents leads to the reduction of metmyoglobin (MbFe3+ ) to myoglobin (MbFe2+ ) [40]. The application of probiotic microorganisms in fermented meat products may signicantly inuence lipid oxidation and color loss in meat products. Most lactobacilli form hydrogen peroxide by oxidizing lactate [15]. Hydrogen peroxide can interfere with the sensory properties of fermented sausages by increasing the rancidity and discoloration of the nal product. The interaction of myoglobin with H2 O2 activates metmyoglobin, which, as a very unstable ferryl myoglobin radical, can transform rapidly into the peroxyl radical form. Ferryl myoglobin has been documented as a

 D. Koozyn-Krajewska, Z.J. Dolatowski / Process Biochemistry 47 (2012) 17611772 Table 2 Examples of potential utilization of probiotic bacteria in meat products. Microbial strain Lactobacillus sake, pediococcus acidilactici Meat product Meat starter cultures Findings L. sake and P. acidilactici in simulated gastrointestinal conditions had the strongest capacity for surviving acidic conditions and 0.30 bile salts, the strains may be regarded as potentially probiotic. Strains seem to suitable for the production of dry sausages, L. rhamnosus E-97800 had the faster growth rate and acidication. It was concluded that NTMS cultures, may be used to increase the safety of Hungarian salami because these cultures gave strong inhibition of both E. coli O111 and List. monocytogenes. Reference Erkkil and Petj [23]

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Lactobacillus rhamnosus GG, LC-705 and E-97800 NTMS cultures, including Lactobacillus acidophilus LAFTIkL10, L. paracasei LAFTIkL26, L. paracasei 5119, Lactobacillus sp. L24 and Bidobacterium lactis LAFTIkB94 Lactobacillus curvatus, Lactobacillus plantarum Potential probiotic 20 strains Lactobacillus (brevis, plantarum, casei, paracsei, pentosus, reuteri, curvatus, sakei, zeae) Potential probiotic L. plantarum/pentosus (MF1291, MF1298, MF1300)

Dry sausage

Erkkil et al. [24]

Hungarian salami

Pidcock et al. [78]

Two types of Greek naturally fermented dry sausages Fermented sausages

Scandinavian-type fermented sausage

1000 strains isolated from Iberian dry fermented sausages (363), and human (337) and pig feces (300) Potential L. fermentum HL57, L. reuteri PL519, L. reuteri PL542 Probiotic Lactobacillus fermentum HL57 Pediococcus acidilactici SP979

Iberian dry-fermented sausages Dry-fermented Iberian sausages Iberian dry-fermented sausages

58% of Lactobacillus curvatus strains and all Lactobacillus plantarum strains isolated from sausages were resistant to 0.3% bile salts and had antimicrobial activity. Lactobacillus strains capable of surviving the pH of the stomach and the environment of the intestine, which make them potential probiotics and may be eventually used as probiotic starter cultures in novel dry fermented sausages manufacture. Strains were able to produce a fermented sausage with similar quality as sausage fermented with the commercial starter culture and were able to survive and establish in an environment similar to the human GIT, inhibit potential pathogenic bacteria in vitro and were considered safe to be used with regard to their antibiotic resistance pattern. Most of these strains are promising probiotic meat culture candidates suitable for Iberian dry fermented sausages. L. fermentum HL57, L. reuteri PL519, and L. reuteri PL542 were identied as a potential probiotic meat starter culture suitable for manufacture of dry-fermented Iberian sausages. Iberian dry-fermented sausages inoculated with L. fermentum HL 57 and P. acidilactici SP979 may be considered as functional products given the counts of these strains found at the end of processing. Probiotic strains of Lactobacillus casei OCK 0900 and OCK 0908, added to the cured pork loins, grew and achieved a count of 107 log cfu/g, and in the samples with the additive of 0.2% glucose, their count was 108 log cfu/g. The results of the experiment showed that probiotic strains had a good ability to survive in dry fermented pork loins and this can be used for dry fermented loin production. L. casei OCK 0900 strain had a good ability to survive in dry-cured loins and it can be used for dry fermented loins production.

Papamanoli et al. [75]

Pennacchia et al. [77]

Klingberg et al. [54]

Ruiz-Moyano et al. [84]

Ruiz-Moyano et al. [85]

Ruiz-Moyano et al. [86]

Lactobacillus casei OCK 0900 and Lactobacillus casei OCK 0908

Dry fermented pork loins

Neffe and [70] Koozyn-Krajewska

Lactobacillus acidophilus Bauer and Lactobacillus casei Bif3 /IV Probiotic L. casei OCK 0900

Dry fermented pork loins

Jaworska et al. [46]

Dry fermented pork loins

et al. [71] Neffe-Skocinska

potential powerful agent which contributes to lipid oxidation in meat products during curing and chilling storage [65]. On the other hand, Fadda et al. [27] stated that certain lactobacilli, such as L. casei, L. plantarum, L. curvatus and L. sakei strains, actively contribute to the hydrolysis of sarcoplasmic proteins, such as myoglobin [3]. Talon et al. [100] found that lactobacilli and staphylococci inhibit the oxidation of unsaturated fatty acids and can reduce Mb(Fe3+ ) to Mb(Fe2+ ) and change meat color from brown or gray to bright red. Probiotic strains have a signicant inuence on the oxidation-reduction potential of meat products. In some cases probiotic strains reveal a synergistic effect of nitrite which inhibits C. botulinum and Listeria monocytogenes [106]. Changes in myoglobin from a ferrous (Fe2+ ) to a ferric (Fe3+ ) state are critical to muscle-based food product appearance, because the consumers use browning as an indicator of spoilage. The presence of metmyoglobin may also have consequences for the oxidative stability especially of unsaturated fatty acids, because this ferric state of myoglobin acts as a pro-oxidant [29,113]. Ruiz-Moyano et al. [86] pointed out that inoculation with a potential probiotic L. fermentum HL57 strain increased the amount of malonaldehyde in Iberian dry-fermented sausages resulting in an unattractive color and taste due to hydrogen peroxide formation. Hydrogen peroxide can react with myoglobin (Fe2+ ) yielding ferryl myoglobin (Fe4+ ) which acts as a pro-oxidant on lipids. Therefore,

a good idea to prevent the oxidative effect of H2 O2 may be to add an antioxidant with the probiotic strain. The oxidative and color stability of meat products depends mostly on the balance of anti- and pro-oxidants. On the other hand, several authors have postulated a strong relationship between cured hem pigments oxidation and lipid oxidation in meat and cured meat products. Free radicals produced during lipid oxidation can oxidize hem pigments to the metmyoglobin form causing discoloration of meat products [51,105]. The results of a study by Wjciak et al. [110] suggest that probiotic strains L. casei OCK 0900 and Lactobacillus paracasei OCK 0919 can be used as ingredients during the curing of pork meat. The addition of these probiotic strains to pork meat samples signicantly reduces pH values not only in meat stufng but also in cooked meat products during whole storage. A sample cured with probiotic bacteria at 4 C for 24 h was more effective in protecting the red color of meat because of its reductive activity (low redox potential value, low TBARS value), delayed lipid oxidation and discoloration over 30 days of storage in comparison to a sample without probiotic strains. Water availability is one of the most important factors affecting microbial growth and survival. In meat, various substances can bind water molecules and thus render them unavailable for microorganisms. Furthermore, solutes, such as salts and sugar, bind water. The

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Table 3 Flow diagram of probiotic dry fermented meat products manufacturing. Step of technological process Pork meat and pork back fat (sausage) Curing of meat (28 days) Manufacturing of meat product Fermentation (24 days) Ripening (21 days) Parameters pH24 <6.2 Hygienically acceptably Meat 2448 h post mortem Temperature 02 C Salt <3% Nitrite <150 mg/kg Nitrate <200 mg/kg Sodium ascorbate <10 mg/kg Sausages, hams, loins Probiotic bacteria 106 jtk/g Glucose <0.8% Spices Temperature 2224 C Relative humidity 8595% Air ow >1 m/s pH 5.45.8 aw 0.950.98 Temperature 1624 C Relative humidity 7585% Air ow <1 m/s pH 4.75.4 aw <0.95 Temperature 46 C pH 4.95.2 aw 0.800.85

Storage and further maturation (till 6 months)

presence of salts and sugars can in effect be considered analogous to a dry environment. In fermented products the dry environment is thus created by a combination of the actual loss of water, e.g. 2550% of the total weight for dry sausages or 1015% for semidry sausages, and binding of water by NaCl and other components. Most bacteria grow only at aw = 0.901.00. The addition of salt subsequently results in reduced water activity of the product which is indeed unfavorable for the growth of probiotics [111]. Staphylococcus aureus is a pathogen most insensitive to water; it grows at aw = 0.86. Since no pathogenic bacteria grow below aw = 0.85, this value has special signicance in the regulations dening low-acid foods. We propose a ow diagram for probiotic meat products, see Table 3. 5. Microbiological, sensory and physicochemical parameters of probiotic meat products 5.1. Survival of probiotic bacteria used in the production of dry fermented meat products The novel role of probiotics used in meat products is primarily in the fermentation process. It is supposed that the meat matrix consisting of meat and fat encapsulates the bacteria and protects them during the critical passage through the gastrointestinal tract. Considering the ability of meat and meat products to shield the probiotic bacteria in the gastrointestinal tract, the use for health purposes of probiotics in the processing of fermented sausages has attracted much attention [111]. Lcke [62] reported that probiotic bacteria survive poorly in fermented meat products. However, as indicated Hammes and Hertel [37] probiotic bacteria could be better adapted at the meat environment than LAB. In a study by Klingberg et al. [54], added lactic acid bacteria, including probiotic strains, persisted in a high count of viable cells during consumption and throughout the storage time of probiotic products. pH changes are also a challenge for the survival of probiotics in fermented meat products. pH reduced from 5.6 to 4.9 after

fermentation limited the survival of probiotics (L. rhamnosus GG and E-97800) in the fermented sausage over the entire fermentation and ripening process [25]. Curing agents such as sodium nitrite are added to the meat batter for preservation. The potential of probiotic bacteria to resist this compound is one of the challenges faced in meat fermentation. Results of several evaluations show that cell viability in a fermented meat environment is strain-dependent. In a study by Arihara et al. [6], L. gasseri was showed to grow better than L. acidophilus, L. crispatus, L. amylovorus, L. gallinarum and L. johnsonii in meat containing 2.5% NaCl and 5 ppm sodium nitrite. Also Jaworska et al. [46] claimed that both the microbiological and sensory quality of fermented pork loin depended on the kind of probiotic strains used for fermentation. Two potentially probiotic strains were used: L. acidophilus Bauer and L. casei Bif3 /IV. After the ripening and storage process, the LAB count in samples with probiotic L. acidophilus Bauer and L. casei Bif3 /IV strains was higher by approximately one logarithmic level than controls (without probiotics). After 21 days of ripening, the highest count of LAB was observed in the sample with L. casei Bif 3/IV with or without 0.2% glucose added (7.95 log cfu/g and 7.84 log cfu/g). The LAB count in loins with L. acidophilus Bauer was 7.92 log cfu/g for samples without glucose and 7.69 log cfu/g for samples with glucose. However, after 180 days of storage, samples with L. casei Bif3 /IV had better LAB survivability than those with L. acidophilus Bauer (7.00 log cfu/g in products without glucose and 7.53 log cfu/g with glucose), which indicated that storage time did not signicantly affect LAB survivability in samples with the probiotic L. casei Bif3 /IV strain. [70] found that probiotic L. casei Neffe and Koozyn-Krajewska strains OCK 0900 and OCK 0908 added to cured pork loins grew and achieved a count of 107 log cfu/g; in the samples with 0.2% glucose added, their count was 108 log cfu/g. Three kinds of loin samples were analyzed in the experiment: two control samples (loins with and without 0.2% glucose), two samples with L. casei OCK 0900 and OCK 0908 probiotic strains added only, and samples with the probiotic strain and 0.2% glucose added. The loin samples prepared in this way ripened for 3 weeks at 16 C. Subsequently, the products were vacuum-packed and stored at 4 C for 6 months. The count of probiotic bacteria in loins stored for 6 months was by 12 log levels lower compared to the products after the ripening process, but the count was still high enough to consider those products to be probiotic. The objective of a subsequent study et al. [71] was to optimize the fermentation by Neffe-Skocinska conditions of pork loins with probiotic bacteria added. The highest LAB growth, including the L. casei OCK 0900 probiotic strain, was found in the pork loins during the 21 day ripening period at 20 C (from 107 to 108 cfu/g) and the lowest LAB growth, including the probiotic strain, in the loin samples aged at 16 C (from 104 to 106 cfu/g). The sensitivity of probiotic bacteria to sodium chloride and sodium nitrite used for the processing of meat can be reduced by the physical treatment of probiotic cells. Arihara and Itoh [6] demonstrated that UV irradiation of L. gasseri generated several mutants resistant to these compounds. Poor survival of probiotic bacteria during meat fermentation can be also overcome by microencapsulation [68]. 5.2. Lipid and protein oxidation in probiotic products The shelf life of fermented meat products is generally not limited by bacterial deterioration, but by physical and chemical spoilage [69]. It is fat stability that is the main restriction on the shelf life of dry fermented meat products [92,98]. Processed products, which are minced, mixed with salt, and heated, expose muscle tissue to oxidative stress responsible for the loss of quality, oxidative avors and loss of hem pigments and vitamins [67,98]. Lipid oxidation can

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also have a negative effect on the nutritional value, and may be responsible for the production of toxic compounds which can trigger metabolic disorders such as mutagenesis and carcinogenesis [33]. Navarro et al. [69] reported that enzymatic hydrolysis during fermentation accelerates lipid peroxidation. In addition, as discussed above, a strong correlation between lipid and myoglobin oxidation, especially in fresh meat, has been documented. Faustman et al. [28] reported that lipid oxidation products, such as 2-heptenal, 2-nonenal and 4-hydroxy-2-nonenal, promote pigment oxidation. However, there is also strong evidence that hem pigments may initiate lipid oxidation [92]. Most publications concerning the incorporation of probiotic bacteria into foods focus only on their survival during manufacture and storage and only a few studies consider the effect of probiotic adjuncts on the oxidative stability of raw meat products. Oxidative processes in probiotic meat products lead to the degradation of lipids and proteins (including pigments) which in turn contributes to the deterioration in color, texture (cross linking binding) and avor. This causes the loss of avor, color and nutritional values and reduces the shelf-life of dry fermented sausages [55,72,91,93,110]. Numerous authors [5,65,110] have found that certain probiotic bacteria of the Bidobacterium and Lactobacillus type affect the fatty acids prole by converting unsaturated acids into Conjugated Linoleic Acid (CLA) through isomerization, hydrogenation and dehydration. Most frequently, linoleic and linolenic acids are subject to these transformations. It is considered that linoleic acid conversion to CLA is a detoxication mechanism used by bacterial cells. By producing health-promoting components, such as CLA, probiotics impact the functional characteristics of food. To extend the shelf life of meat products, certain antioxidants are added. Sodium ascorbate is the most popular antioxidant added to dry-fermented sausages in meat industry. Nevertheless, consumers prefer natural antioxidants, such as rosemary, sage or green tea extract, not only due to their health-promoting properties but also due to greater antioxidizing properties as compared to synthetic antioxidants [1,14]. Studies of the effects of the potentially probiotic L. acidophilus CH-2 strain and green tea infusion on the oxidative stability of ripening pork loin showed that adding the L. acidophilus CH2 strain, glucose and green tea infusion increased the oxidative stability of the meat product by lowering the TBARS value in comparison with the control sample, as well as improving product redness [46]. Authors also observed better color stability during exposure to light. Skwarek and Dolatowski [93] examined raw ripening hams with added probiotic bacteria and green tea infusion; they observed lower TBARS values (0.385 mg MDA/kg and 0.97 mg MDA/kg, respectively) and lower levels of oxidized myoglobin (46.3% and 43.35%, respectively) in comparison with the control sample (without probiotic strains). However, the authors also found an increase in the oxidative index (0.524 mg MDA/kg) and metmyoglobin (72.5%) and a loss of redness in samples with an added probiotic L. casei OCK 0900 strain which were not treated with antioxidants. The increase in the TBARS value and metmyoglobin was higher in potentially probiotic products compared to the control product (without probiotic strains). Primary and secondary products of lipid autoxidation reacting with proteins are less reactive than free radicals; however, they can move within a biological material [16,19,26,107]. Aldehydes formed in a lipid peroxidation reaction react mainly with triol and amine groups in proteins, amino sugars and nitrogen bases in nucleic acids. Recombination of protein free radicals with cysteine or tyrosine residues leads to the formation of protein dimers linked by cysteine or bis-tyrosine bridges [30]. Losses of exogenous amino acids play an essential role in nutrition, because they reduce the nutritional value of protein in

products. The amino acids most sensitive to oxidation are heterocyclic amino acids. Due to their structure, tryptophan, histidine, proline, lysine, cysteine, methionine and tyrosine are prone to oxidation and a hydrogen atom is eliminated from OH-, S- or Ncontaining groups [10,18,45,49,89]. Our studies of the effect of L. casei OCK 0900 on product oxidative transformations show that when probiotic bacteria are added, the oxidation of product components increases during ripening and the initial storage period [110]. On further storage, oxidation processes stabilize. We hypothesize that probiotic bacteria lead to protein degradation to yield proteins with antioxidative properties which stabilize oxidation processes in the product. 5.3. Sensory properties of probiotic fermented meat products The inuence of microorganisms on the sensory quality of products with probiotic properties raises increasingly more interest; however, research reports on meat products are quite scarce. Products fermented with intestinal bacteria taste different than those manufactured in traditional ways: they are mildly acidic and usually with an unspecied aroma. Such sensory properties may be unacceptable to consumers and, therefore, the avor is frequently corrected through the addition of herbs, aromas, sweeteners and by increasing dry mass content. In the latest fermented products, an addition of appropriate, so-called prebiotic saccharides is proposed, such as lactulose, fructo-oligosaccharides or galacto-oligosaccharides, which selectively stimulate the growth of bidobacteria in the alimentary tract [19,43]. In 1998, Sameshima [43] discovered that the characteristic avor and aroma of raw sausages with added experimental probiotic bacteria, which were able to control the growth of the pathogenic microbiota, did not differ at all from sausages in which commercial strains were used. Further, Typpnen et al. [104] found that lactic acid bacteria added to raw sausages act as starter cultures and probiotics at the same time. In the study by Klingberg et al. [54] on sausages fermented by various Lactobacillus strains almost identical avor proles were obtained compared with samples produced by the commercial starter culture. In the reports by Jaworska et al. [46] differences, although not signicant, were observed in the acidic taste. It was claimed that the sensory quality of fermented pork loin depended on the variety of probiotic strains used for fermentation: L. acidophilus Bauer or L. casei Bif3 /IV. The highest sensory quality after ripening and chilling storage was observed in samples produced with L. acidophilus Bauer with 0.2% glucose added. The storage process had a strong impact on the decrease in the overall sensory quality, but to a different extent in the tested samples. The highest sensory quality after storage was observed in fermented pork loin samples produced with L. acidophilus Bauer with 0.2% glucose added. et al. [71], the inuence of ripening temIn Neffe-Skocinska perature on sensory quality was studied. Product samples ripened at 16 and 20 C had the highest overall quality. With respect to both the microbiological and sensory quality, the optimal ripening conditions of the dry raw loins with probiotic bacteria added in laboratory conditions were: temperature of 20 C and time of 21 days. In the case of industrial production the temperature of 16 C should be rather used for safety reasons. Zdolec et al. [112] showed that the addition of a probiotic L. sakei culture to fermented sausages did not have a negative impact on the sensory properties of the sausages. Furthermore, improvement in sensory parameters, mainly acidity, juiciness and tenderness was observed. Erkkila et al. [24,25] also reported that the (potential) probiotic L. rhamnosus GG, L. rhamnosus LC-705, L. rhamnosus E97800 and L. plantarum E-98098 strains did not negatively affect the technological or sensory properties of the end product (sausage),

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with a (minor) exception of L. rhamnosus LC-705. When intestinal isolates of L. paracasei L26 and Bidobacterium lactis B94 were used in combination with a traditional meat starter culture, no negative impact on the sensory properties of the product was noted [78]. 6. Probiotic meat products in human nutrition benets and risks 6.1. Human studies using probiotic fermented meats Probiotic bacteria are used as additives in food to provide a wide variety of health benets, including improving the function of the immune system and prevention of cancer, atherosclerosis and coronary disease [84,103]. To study probiotic properties of lactic acid bacteria, such as L. casei/paracasei, L. rhamnosus and L. sakei, the survival rate in gastric and intestinal juice was examined by the in vitro adhesion to intestinal cells, production of organic acids and deactivation of pathogenic microorganisms. Besides, it was proved that several L. plantarum strains display greater adhesion to intestinal mucosa than L. brevis and L. paracasei (strain groups isolated from sausages). However, the fact that all properties are seen does not necessarily result in the intensication of health-benecial properties, which principally results from the fact that the studies conducted are incomplete. Human studies using probiotic fermented meats have been very scarce until recently, but they are crucial in the validation process required for the conrmation of the potential function of probiotic fermented products. As an example, the effect of the daily consumption of 50 g of probiotic sausages containing L. paracasei LTH 2579 on immunity and blood serum lipids was investigated in healthy volunteers (10 women and 10 men) over 5 weeks. Modulation of various aspects of host immunity was observed, but there was no signicant inuence on the serum concentration of different cholesterol fractions and triacylglycerides. In fecal samples, a statistically signicant increase in the numbers of L. paracasei LTH 2579 was observed for some but not all volunteers. It was concluded that bacterial adhesion to enterocytes, fecal colonization or both seem to be an important criterion for immunomodulation. Because L. paracasei can transiently colonize the intestine, authors hypothesize that this strain has contact with immune cells and that immunomodulation can thus occur [44]. Consumption of probiotic meats can be benecial also because of the nutritional value of such products. According to JimnezColmenero et al. [47] dry-cured ham has a well-balanced nutritional prole and is suitable in appropriate amounts for consumption (even 23 times per week in healthy individuals) as part of a balanced diet. While a certain amount of ham consumed can produce benecial effects with regard to cardiovascular disorders, more scientic evidence is needed to assess the function ham can have in relation to health. The major objections in this respect arise from aspects associated with the quality of the fat and the presence of sodium. Ham contains large quantities of monounsaturated and polyunsaturated fatty acids, but because the proportion of n-6 PUFA is relatively high, the n-6/n-3 ratio is higher than recommended. 6.2. Safety of probiotic meat products The application of probiotic microorganisms to fermented meat products may be the most effective technology for controlling pathogen growth and survival as a result of production of bacteriocins and other low molecular mass antimicrobials that are inhibitory to undesirable organisms. For instance, Lactobacillus reuteri ATCC 55730 and Bidobacterium longum ATCC 15708 increased the inactivation of Escherichia coli O157:H7 during sausage manufacturing [68]. L. rhamnosus FERM P-15120 and

L. paracasei subsp. Paracasei FERM P-15121 inhibited the growth and enterotoxin production of S. aureus to the same extent as a commercial L. sakei starter culture [88]. In contrast, L. acidophilus FERM P-15119 could not satisfactorily decrease S. aureus numbers, indicating the importance of careful strain selection with respect to both health-promoting and food safety properties. Lactic acid bacteria (LAB) in foods have a long history of safe use. Members of the genera Lactococcus and Lactobacillus are most commonly given the generally recognized-as-safe (GRAS) status, while members of the genera Streptococcus and Enterococcus and some other LAB genera contain certain opportunistic pathogens. Probiotics may be responsible for four types of side effects: systemic infections, risk of deleterious metabolic activity, risk of adjuvant side effects, risk of immunomodulation and risk of gene transfer [56]. Based on literature data on documented cases of infections caused by probiotics and epidemiological data, Koozyn-Krajewska et al. [56] assessed the risk involved in meat probiotic products. The risk factors related to both consumer safety and probiotic meat product safety were identied. Health condition, physiological condition and consumer age were included in the group of consumer risk factors. Microbiological hazards and other hazards (e.g. connected with animal health, animal tracing and the HACCP system in meat production, operations and technologies of meat processing) were identied as meat product safety factors. Microbial hazards are related to the presence not only of pathogens in meat products and their interactions with probiotic bacteria, simply to the presence but also of probiotic bacteria in food (so-called side effects). It was concluded that Enterococcus species are the most serious risk for consumers connected with probiotic meat products consumption. The host factors appear to play a key role in the establishment of an LAB infection and contact of those at risk with these bacteria should be minimized.

6.3. Biogenic amines in probiotic meat products Biogenic amines are organic bases of low molecular weight and with biological activity. They can be formed and degraded as a result of normal metabolic activity in animals, plants and microorganisms, and are usually produced by the decarboxylation of free amino acids or by the amination and transamination of aldehydes and ketones. The removal of the -carboxyl group from a precursor amino acid leads to a corresponding biogenic amine [10,12,63,95]. Tyramine and histamine are principal examples of protein decarboxylation found in the highest quantities in meat. During fermentation or spoilage, protein breakdown products, peptides and amino acids are precursors for amine formation [10,97,108]. Meat and meat products have repetitively been reported to contain biogenic amines [12,21,31,38,49,63,83,94]. The only amines present in signicant levels in fresh meat are spermidine and spermine [38]. High spermine contents, usually between 20 and 60 mg/kg, are usual in meat and meat products of warmblooded animals. Spermidine levels in meat rarely exceed 10 mg/kg [48]. Fermented meat products and especially dry fermented sausages are one of the most common sources of these microbial metabolites. During fermentation, maturation and storage of dry fermented sausages suitable environmental conditions occur that favor the activity of microorganisms containing decarboxylase enzymes, and thus the accumulation of biogenic amines, mainly tyramine, histamine, phenylethylamine, tryptamine, putrescine and cadaverine, may occur [2,50,99]. Protein decomposition occurs with the participation of microbial enzymes and often results in deamination and decarboxylation, which is additionally supported by the acidic environment.

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Amine production has been recognized as a defense microbial mechanism against an acidic environment [50,99]. Tkachenko et al. [101] suggested an interesting hypothesis on the physiological role of biogenic amines in microorganisms. Amino acid decarboxylases are enzymes found in many microorganisms which may be either naturally present in food products or may be introduced by contamination before, during or after food processing. They have been found in species of the genera Bacillus, Clostridium, Pseudomonas, Photobacterium, as well as in genera of the family Enterobacteriaceae, such as Citrobacter, Klebsiella, Escherichia, Proteus, Salmonella and Shigella, and Micrococcaceae such as Staphylococcus, Micrococcus and Kocuria. Furthermore, many lactic acid bacteria belonging to the genera Lactobacillus, Enterococcus, Carnobacterium, Pediococcus, Lactococcus and Leuconostoc are able to decarboxylate one or more amino acids [31,50,99]. There is much evidence that in fermented meat products the contaminating microora rather than the starter cultures is responsible for the generation of an increasing biogenic amine level [10]. The meat raw material is the natural source of substrates from which biogenic amines are produced. It is also the largest component of the matrix in which decarboxylation reactions take place and any conditions that alter its nature and characteristics will inuence the formation of biogenic amines [83]. Biogenic amine accumulation in dry fermented sausages depends mainly on the hygienic quality of the raw meat material and the processing plant, since most contaminant bacteria, such as enterobacteria and pseudomonads, can produce amines [83,99]. The relevance of Gram-negative bacteria for biogenic amine accumulation during sausage fermentation has been demonstrated previously [12,13]. Although found at a lower frequency than in such contaminants, 1020% of fermentative microbiota isolates, including lactic acid bacteria and Gram-positive catalase-positive cocci, have also been reported to possess aminogenic potential. More specically, particular strains of species that are usually involved in the sausage fermentation process, such as L. brevis, L. curvatus, Enterococcus faecalis, Enterococcus faecium and Staphylococcus carnosus, are able to produce one or more biogenic amines [13,59,97]. Several authors [21,63] observed signicant differences in biogenic amine content in fermented meat products depending on raw material hygienic quality. However, the same raw material can lead to very different amine levels in nal products depending on the presence of decarboxylating microorganisms, either derived from environmental contamination or from starter cultures, and the conditions supporting their growth vary depending on several extrinsic and intrinsic factors during the manufacturing process (such as pH, redox potential, temperature, NaCl, size of the sausage, hygienic conditions of manufacturing practices and effect of starter cultures) [32,59]. The biogenic amines found in meat products are currently considered a potential cause of headaches, circulation disorders and poisoning. When the fermentation process is managed properly, the concentration of biogenic amines is low and consumption of raw cured meat products cannot lead to food poisoning. Undesired accumulation of biogenic amines in meat products requires the availability of precursors (i.e. amino acids), the presence of microorganisms with amino acid decarboxylases, either derived from environmental contamination or from an added starter culture and favorable conditions that allow bacterial growth, decarboxylase synthesis and decarboxylase activity [2,10,45,50,99]. Certain amines, such as tyramine, cadaverine, and putrescine, can be formed during the storage of meat [31,38]. Tyramine concentrations in stored beef were found to be the highest on the meat surface; therefore, this can be effectively reduced by washing [49,76].

Additionally, the microorganisms responsible for the fermentation process may contribute to biogenic amine accumulation [13,59]. The non-protein nitrogen fraction which increases during fermentation includes the presence of free amino acids, precursors of biogenic amines. The major protease activity is derived from endogenous meat enzymes. Proteolysis is favored by the denaturation of proteins as a consequence of acidity increase, dehydration and action of sodium chloride [21,99]. Fermented products with comparable microbiological proles may differ in their biogenic amine content, indicating that the production of such compounds depends on a complex interaction of factors [99]. In a study by Stadnik and Dolatowski [95], biogenic amine changes during ageing of dry-cured pork loins inoculated with probiotics were analyzed in 4-, 8- and 16-month-old samples. The comparison of biogenic amine contents in samples of dry-cured pork loins inoculated with probiotic L. casei OCK 0900 showed great variability. Histamine and spermidine were not detected; spermine was present at very low levels, while cadaverine and tryptamine were the most abundant biogenic amines. Their concentrations were in agreement with previously reported results for other dry cured meat products and below the suggested toxic limits. The dry cured loins were characterized by gradually decreasing water activity and rapid reduction in pH compared to raw meat; this may be regarded as a mild background suitable for the generation of biogenic amines [95]. Bacterial amino acid decarboxylases usually have an acidic pH optimum. Reduced pH results in increased decarboxylase activity in bacteria. In these conditions bacteria produce more decarboxylases as part of their protective mechanism. However, rapid and large pH reduction is known to reduce the growth of amine-positive microorganisms [63]. Amine formation in bacteria is decisively inuenced by temperature. Temperatures between 20 C and 37 C are optimal for the growth of most decarboxylase-containing bacteria; reduced temperature stops their growth [50]. According to Suzzi and Gardini [99], biogenic amine accumulation decreases markedly with the increase of NaCl concentration, while proteolytic activity is higher for the intermediate concentration of salt; this suggests that there is not necessarily a correlation between these two variables. Karovi cov and Kohajdov [50] reported that the presence of sodium chloride activates tyrosine decarboxylase activity and inhibits histidine decarboxylase activity. However, some strains of L. curvatus used as starter cultures form up to four different biogenic amines and should not be used. Strains of L. sakei do not have this potential [11,55,108]. As far as possible negative aspects of the consumption of meat and meat products are concerned, there is a growing interest in polyamines which have no adverse health effects, but have been described as potential precursors of stable carcinogenic Nnitrosamines and to enhance the growth of chemically induced aberrant crypt foci in the intestine. The formation of N-nitroso compounds constitutes an additional toxicological risk associated with biogenic amines, especially in meat products that contain nitrite and nitrate salts as curing agents [50,89].

7. Conclusions and future trends Meat products provide an adequate environment for the growth of probiotic microbiota, and recently there have been attempts to use probiotic bacteria in the production of fermented meat products. However, the production of probiotic meat products requires overcoming certain technological limitations, such as the native microora of meat, a need to use additives such as nitrites and salt and also low water activity and low content or absence of natural sugars. Probiotic bacteria strains that can be used in the

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D. Koozyn-Krajewska, Z.J. Dolatowski / Process Biochemistry 47 (2012) 17611772 [11] Bover-Cid S, Izquierdo-Pulido M, Vidal-Carou MC. Effectiveness of a Lactobacillus sakei starter culture in the reduction of biogenic amine accumulation as a function of the raw material quality. J Food Prot 2001;64(3):36773. [12] Bover-Cid S, Miguelez-Arrizado MJ, Latorre Moratalla LL, Vidal Carou MC. Freezing of meat raw materials affects tyramine and diamine accumulation in spontaneously fermented sausages. Meat Sci 2006;72:628. [13] Bover-Cid S, Hugas M, Izquierdo-Pulido M, Vidal-Carou MC. Amino acid decarboxylase activity of bacteria isolated from fermented pork sausages. Int J Food Microbiol 2001;66(3):1859. [14] Bozkurt H. Utilization of natural antioxidants: green tea extract and Thymbra spicata oil in Turkish dry-fermented sausage. Meat Sci 2006;73:44250. [15] Caplice E, Fitzgerald GF. Food fermentations: role of microorganisms in food production and preservation. Int J Food Microbiol 1999;50(12):13149. [16] Carlsen CU, Mller JKS, Skibsted LH. Heme-iron in lipid oxidation. Coord Chem Rev 2005;249:48598. [17] Cassens R. Use of sodium nitrite in cured meats today. Food Technol 1995;7:7280. [18] Chelh I, Gatellier P, Sant-Lhoutellier V. Characterisation of uorescent Schiff bases formed during oxidation of pig myobrils. Meat Sci 2007;76:2105. [19] Collins MD, Gibson GR. Probiotics, prebiotics, and synbiotics: approaches for modulating the microbial ecology of the gut. Am J Clin Nutr 1999;69:1052S7S. [20] De Vuyst L, Falony G, Leroy F. Probiotics in fermented sausages. Meat Sci 2008;80:758. [21] Eerola S, Roig Sagus A-X, Lilleberg L, Aalto H. Biogenic amines in dry sausages during shelf-life storage. Z Lebensm Unters Forsch 1997;205(A):3515. [22] Erkkil S. Bioprotective and probiotic meat starter cultures for the fermentation of dry sausages. Academic dissertation. Helsinki: University of Helsinki; 2001. [23] Erkkil S, Petj E. Screening of commercial meat starter cultures at low pH and in the presence of bile salts for potential probiotic use. Meat Sci 2000;55:297300. [24] Erkkil S, Petj E, Eerola S, Lilleberg L, Mattila-Sandholm T, Suihko ML. Flavour proles of dry sausages fermented by selected novel meat starter cultures. Meat Sci 2001;58:1116. [25] Erkkila S, Suihko ML, Eerola S, Petaja E, Mattila-Sandholm T. Dry fermented sausages by Lactobacillus rhamnosus strains. Int J Food Microbiol 2001;64:20510. [26] Estvez M, Cava R. Effectiveness of rosemary essential oil as an inhibitor of lipid and protein oxidation: contradictory effects in different types of frankfurters. Meat Sci 2006;72:34855. [27] Fadda S, Oliver G, Vignolo G. Protein degradation by Lactobacillus plantarum and Lactobacillus casei in a sausage model system. J Food Sci 2002;67(3):117983. [28] Faustman C, Liebler DC, McClure TD, Sun Q. alpha,beta-Unsaturated aldehydes accelerate oxymyoglobin oxidation. J Agric Food Chem 1999;47:31404. [29] Faustman C, Sun Q, Mancini R, Suman SP. Myoglobin and lipid oxidation interactions: mechanistic bases and control. Meat Sci 2010;86:8694. [30] Fellenberg MA, Speisky H. Antioxidants: their effects on broiler oxidative stress and its meat oxidative stability. World Poultry Sci J 2006;62:5370. [31] Galgano F, Favati F, Bonadio M, Lorusso V, Romano P. Role of biogenic amines as index of freshness in beef meat packed with different biopolymeric materials. Food Res Int 2009;42:114752. [32] Gardini F, Martuscelli M, Caruso MC, Galgano F, Crudele MA, Favati F, et al. Effects of pH, temperature and NaCl concentration on the growth, kinetics, proteolytic activity and biogenic amine production of Enterococcus faecalis. Int J Food Microbiol 2001;64:10517. [33] Ghiretti GP, Zanardi E, Novelli E, Campanini G, Dazzi G, Madarena G, et al. Comparative evaluation of some antioxidants in salami Milano and Mortadella production. Meat Sci 1997;47(1/2):16776. [34] Hammes WP. Metabolism of nitrate in fermented meats: the characteristic feature of a specic group of fermented foods. Food Microbiol 2012;29: 1516. [35] Hammes WP, Vogel RF. The genus Lactobacillus. In: Wood BJB, Holzapfel WH, editors. The lactic acid bacteria, vol. 2. London: Chapman and Hall; 1995. p. 1954. [36] Hammes WP, Hertel C. Selection and improvement of lactic acid bacteria used in meat and sausage fermentation. Lait 1996;76:15968. [37] Hammes WP, Hertel C. New developments in meat starter cultures. Meat Sci 1998;49:S12538. [38] Hernndez-Jover T, Izquierdo-Pulido M, Veciana-Nogus MT, Marin-Font A, Vidal-Carou MC. Biogenic amine and polyamine contents in meat and meat products. J Agric Food Chem 1997;45:2098102. [39] Holzapfel WH. Introduction to prebiotics and probiotics. In: Goktepe I, Juneja VK, Ahmedna M, editors. Probiotics in food safety and human health. Boca Raton: CRC Press; 2006. p. 134. [40] Honikel KO. The use and control of nitrate and nitrite for the processing of meat products. Meat Sci 2008;78:6876. [41] Hugas M, Monfort JM. Bacterial starter cultures for meat fermentation. Food Chem 1997;59(4):54754. [42] Incze K. Dry fermented sausages. Meat Sci 1998;49:S16977. [43] Incze K. Fermentierte Fleischprodukte. Fleischwirtschaft 2002;4:1127. [44] Jahreis G, Vogelsang H, Kiessling G, Schubert R, Bunte C, Hammes WP. Inuence of probiotic sausage (Lactobacillus paracasei) on blood lipids and immunological parameters of healthy volunteers. Food Res Int 2002;35:1338.

manufacturing of dry fermented meat products should be capable of surviving in conditions found in fermented products; furthermore, they should dominate other microorganisms found in the nished product. Moreover, the product should maintain its sensory characteristics. The quality of the nal product (e.g. ham, loin, sausage) is closely related to ripening and storage. The process favors the growth of probiotic microorganisms, which inuences the sensorial and nutritional qualities, safety and other characteristics of this type of meat products. Probiotic strains should be present in the product in the number at least 106 /g to have an impact on the health of consumers gut ora. Probiotics in meat may exert their benets by way of several mechanisms, and more research is required to identify the strains and combinations of probiotic bacteria strains that produce the most health benets. There is only little scientic evidence published, related to studies of the effect of probiotic meat products on human health. The therapeutic effect of consuming food with probiotic bacteria is a factor contributing to the development of research on this new segment of functional food. Biogenic amines may be a risk factor connected with probiotic meat consumption. Not only the inability of the culture to form biogenic amines but also its ability to grow well at temperatures intended for product processing and competitiveness in suppressing the growth of wild amine producing microora should be taken into consideration in the selection of starter cultures. Biogenic amine accumulation in dry fermented sausages depends mainly on the hygienic quality of the raw meat material and the processing plant. However, the same raw material can lead to very different amine levels in nal products depending on the presence of decarboxylating microorganisms and the conditions supporting their growth vary depending on several extrinsic and intrinsic factors during the manufacturing process. As a tendency is observed that consumers are to a greater extent oriented toward functional products and their increasing role in human nutrition, we expect that in future, probiotic meat products will become an important part of the meat processing industry. They bridge the existing gap in the eld of probiotic products of animal origin, now dominated by dairy products. However, it is of utmost importance to ensure microbiological and chemical safety as well as sensory acceptability of new products.

References
[1] Aguirrezbal MM, Mateo J, Dominguez MC, Zumalacrregui JM. The effect of paprika, garlic and salt on rancidity in dry sausages. Meat Sci 2000;54: 778. [2] Alberto MR, Arena ME, Manca de Nadra MC. A comparative survey of two analytical methods for identication and quantication of biogenic amines. Food Control 2002;13:1259. [3] Ammor MS, Mayo B. Selection criteria for lactic acid bacteria to be used as functional starter cultures in dry sausage production: an update. Meat Sci 2007;76:13846. [4] Andersen L. Fermented dry sausages produced with the admixture of probiotic cultures. In: Proceedings of 44th ICoMST. 1998. p. 8267. [5] Arihara K. Strategies for designing novel functional meat products. Meat Sci 2006;74:21929. [6] Arihara K, Itoh M. UV-induced Lactobacillus gasseri mutants resisting sodium chloride and sodium nitrite for meat fermentation. Int J Food Microbiol 2000;56:22730. [7] Bekhit AED, Faustman C. Metmyoglobin reducing activity. Meat Sci 2005;71: 40739. [8] Benito MJ, Martn A, Aranda E, Prez-Nevado F, Ruiz-Moyano S, Crdoba MG. Characterization and selection of autochthonous lactic acid bacteria isolated from traditional Iberian dry-fermented salchichn and chorizo sausages. J Food Sci 2007;72:M193201. [9] Benito MJ, Serradilla MJ, Ruiz-Moyano S, Martn A, Prez-Nevado F, Crdoba MG. Rapid differentiation of lactic acid bacteria from autochthonous fermentation of Iberian dry-fermented sausages. Meat Sci 2008;80(3):65661. [10] Bodmer S, Imark C, Kneubhl M. Biogenic amines in foods: histamine and food processing. Inamm Res 1999;48:296300.

 D. Koozyn-Krajewska, Z.J. Dolatowski / Process Biochemistry 47 (2012) 17611772 [45] Jansen SC, van Dusseldorp M, Bottema KC, Dubois AEJ. Intolerance to dietary biogenic amines: a review. Ann Allergy Asthma Immunol 2003;91:23341. D, Dolatowski Z. Survival during stor[46] Jaworska D, Neffe K, Koozyn-Krajewska age and sensory effect of potential probiotic lactic acid bacteria Lactobacillus acidophilus Bauer and Lactobacillus casei Bif3 /IV in dry fermented pork loins. Int J Food Sci Technol 2011;46:24917. [47] Jimnez-Colmenero F, Ventanas J, Toldr F. Nutritional composition of drycured ham and its role in a healthy diet. Meat Sci 2010;84:58593. c P, Krausov P. A review of dietary polyamines: formation, implications [48] Kala for growth and health and occurrence in foods. Food Chem 2005;90:21930. [49] Kaniou I, Samouris G, Mouratidou T, Eleftheriadou A, Zantopoulos N. Determination of biogenic amines in fresh unpacked and vacuum-packed beef during storage at 4 C. Food Chem 2001;74:5159. [50] Karovi cov J, Kohajdov Z. Biogenic amines in food. Chem Pap 2005;59(1):709. [51] Karwowska M, Dolatowski ZJ. An assessment of buckwheat and oat preparations in the production of comminuted meat products. Roczniki Instytutu i Tuszczowego, vol. XLV/1. Warsaw: Instytut Przemysu Przemysu Miesnego i Tuszczowego; 2007. p. 18190. Miesnego [52] Kedia G, Wang R, Patel H, Pandiella SS. Use of cereal-based substrates with potential probiotic properties. Process Biochem 2007;42:6570. [53] Khan MI, Arshad MS, Anjum FM, Sameen A, Rehman A, Gill WT. Meat as a functional food with special reference to probiotic sausages. Food Res Int 2011;44:312533. [54] Klingberg TD, Axelsson L, Naterstad K, Elsser D, Budde BB. Identication of potential probiotic starter cultures for Scandinavian-type fermented sausages. Int J Food Microbiol 2005;105:41931. [55] Koozyn-Krajewska D, Dolatowski ZJ. Probiotics in fermented meat products. Acta Sci Pol Technol Aliment 2009;8(2):6176. [56] Koozyn-Krajewska D, Dolatowski ZJ, Zielinska D. Risk assessment of probiotic use particularly in meat products a review. Fleischwirtschaft 2011;4:618. [57] Krckel L. Use of probiotic bacteria in meat products. Fleischwirtschaft 2006;86:10913. [58] Kun S, Rezessy-Szabo JM, Nguyen QD, Hoschke A. Changes of microbial population and some components in Carnot juice during fermentation with selected Bidobacterium strains. Process Biochem 2008;43:81621. [59] Latorre-Moratalla ML, Bover-Cid S, Talon R, Garriga M, Zanardi E, Ianieri A, et al. Strategies to reduce biogenic amine accumulation in traditional sausage manufacturing. LWT-Food Sci Technol 2010;43:205. [60] Leroy F, Verluyten J, De Vuyst L. Functional meat starter cultures for improved sausage fermentation. Int J Food Microbiol 2006;106:27085. [61] Leroy S, Giammarinaro P, Chacornac JP, Lebert I, Talon R. Biodiversity of indigenous staphylococci of naturally fermented dry sausages and manufacturing environments of small-scale processing units. Food Microbiol 2010;27(2):294301. [62] Lcke FK. Utilization of microbes to process and preserve meat. Meat Sci 2000;56:10515. [63] Maijala R, Nurmi E, Fischer A. Inuence of processing temperature on the formation of biogenic amines in dry sausages. Meat Sci 1995;39:922. [64] Martn L, Crdoba JJ, Ventanas J, Antequera T. Changes in intramuscular lipids during ripening of Iberia dry-cured ham. Meat Sci 1999;51:12934. [65] Min B, Ahn DU. Mechanism of lipid oxidation in meat and meat products a review. Food Sci Biotechnol 2005;14:15263. [66] Morita H, Sakata R, Nagata Y. Nitric oxide complex of iron (II) myoglobin converted from metmyoglobin by Staphylococcus xylosus. J Food Sci 1998;63(2):3525. [67] Morrissey PA, Sheehy PJA, Galvin K, Kerry JP, Buckley DJ. Lipid stability in meat and meat products. Meat Sci 1998;49:73. [68] Muthukumarasamy P, Holley R. Microbiological and sensory quality of dry fermented sausages containing alginate-microencapsulated Lactobacillus reuteri. Int J Food Microbiol 2006;111:1649. [69] Navarro JL, Nadal MI, Izquierdo L, Flores J. Lipolysis in dry cured sausages as affected by processing conditions. Meat Sci 1997;45:1618. [70] Neffe K, Koozyn-Krajewska D. Potential uses of probiotic bacteria in ripening Technol Ja 2010;5(72):16777. meat products. Zywn-Nauk K, Gierejkiewicz M, Koozyn-Krajewska D. Optimization of [71] Neffe-Skocinska fermentation conditions for dry-aged sirloins with probiotic bacteria added. Zywn-Nauk Technol Ja 2011;6(79):3646. K, Jachacz L, Jaworska D, Koozyn-Krajewska D, Dolatowski [72] Neffe-Skocinska ZJ. Effect of probiotic bacteria Lactobacillus casei ock 0900 and green tea extract addition on dry fermented pork loins quality; 2012. Unpublished results. zewska [73] Nowak A, Sli K, Libudzisz Z. Probotics history and mechanisms of Technol Ja 2010;71:519. their effect. Zywn-Nauk [74] Palacios MC, Sanz Y, Haros M, Rosell CM. Application of Bidobacterium strains to the breadmaking process. Process Biochem 2006;41:243440. [75] Papamanoli E, Tzanetakis N, Litopoulou-Tzanetaki E, Kotzekidou P. Characterization of lactic acid bacteria isolated from a Greek dry-fermented sausage in respect of their technological and probiotic properties. Meat Sci 2003;65:85967. [76] Paulsen P, Hagen U, Bauer F. Changes in biogenic amine contents, non-protein nitrogen and crude protein during curing and thermal processing of M. longissimus, pars lumborum of pork. Eur Food Res Technol 2006;223:6038. [77] Pennacchia C, Ercolini D, Blaiotta G, Pepe O, Mauriello G, Villani F. Selection of Lactobacillus strains from fermented sausages for their potential use as probiotics. Meat Sci 2004;67:30917.

1771

[78] Pidcock K, Heard GM, Henriksson A. Application of nontraditional meat starter cultures in production of Hulgarian salami. Int J Food Microbiol 2002;76:7581. [79] Probiotics in food. Health and nutritional properties and guidelines for evaluation. FAO Food and Nutrition. Paper 85, Rome; 2006. [80] Rebucci R, Sangalli L, Fava M, Bersani C, Cantoni C, Baldi A. Evaluation of functional aspects in Lactobacillus strains isolated from dry fermented sausages. J Food Qual 2007;30:187201. [81] Roberfroid MB. Prebiotics and probiotics: are they functional foods. Am J Clin Nutr 2000;7(Suppl.):1682S7S. [82] Rozada-Sanchez R, Sattur AP, Thomas K, Pandiella SS. Evaluation of Bidobacterium spp. for the production of a potentially probiotic malt-based beverage. Process Biochem 2008;43:84854. [83] Ruiz-Capillas C, Jimnez-Colmenero F. Biogenic amines in meat and meat products. Crit Rev Food Sci Nutr 2004;44:48999. [84] Ruiz-Moyano S, Martn A, Benito MJ, Nevado FP, Crdoba MdeG. Screening of lactic acid bacteria and bidobacteria for potential probiotic use in Iberian dry fermented sausages. Meat Sci 2008;80:71521. [85] Ruiz-Moyano S, Martn A, Benito MJ, Hernndez A, Casquete R, Serradilla MJ, et al. Safety and functional aspects of pre-selected lactobacilli for probiotic use in Iberian dry-fermented sausages. Meat Sci 2009;83:4607. [86] Ruiz-Moyano S, Martn A, Benito MJ, Hernndez A, Casquete R, Crdoba MdeG. Application of Lactobacillus fermentum HL57 and Pediococcus acidilactici SP979 as potential probiotics in the manufacture of traditional Iberian dry-fermented sausages. Food Microbiol 2011;28:83947. [87] Salminen S, von Wright A, Morelli L, Marteau P, Brassart D, de Vos WM, et al. Demonstration of safety of probiotics a review. Int J Food Microbiol 1998;44:93106. [88] Sameshima T, Magome C, Takeshita K, Arihara K, Itoh M, Kondo Y. Effect of intestinal Lactobacillus starter cultures on the behaviour of Staphylococcus aureus in fermented sausage. Int J Food Microbiol 1998;41:17. [89] Santos SMH. Biogenic amines: their importance in foods. Int J Food Microbiol 1996;29:21331. [90] Schillinger U, Geisen R, Holzapfel WH. Potential of antagonistic microorganisms and bacteriocins for the biological preservation of foods. Trends Food Sci Technol 1996;7:15864. [91] Singh VP, Pathak V, Verma AK. Fermented meat products: organoleptic qualities and biogenic amines a review. Am J Food Technol 2012;7: 27888. [92] Skibsted LH. Chemical changes in meat and meat products during storage, transportation and retail display theoretical considerations. In: Taylor SA, Raimundo A, Severini M, Smulders FJM, editors. Meat quality and meat packaging. Utrecht, The Netherlands: ECCEAMST; 1996. p. 16981. [93] Skwarek M, Dolatowski ZJ. Physicochemical properties of raw ripening hams with the addition of probiotic. Acta Sci Pol Biotechnol 2010;9(1): 1524. [94] Stadnik J, Dolatowski ZJ. Biogenic amines in meat and fermented meat products. Acta Sci Pol Technol Aliment 2010;9(3):25163. [95] Stadnik J, Dolatowski ZJ. Biogenic amines content during extended ageing of dry-cured pork loins inoculated with probiotics. Meat Sci 2012;91:3747. [96] Stanton C, Ross RP, Fitzgerald GF, Van Sinderen D. Fermented functional foods based on probiotics and their biogenic metabolities. Curr Opin Biotechnol 2005;16:198203. [97] Straub B, Schollenberger M, Kicherer M, Luckas B, Hammes WP. Extraction and determination of biogenic amines in fermented sausages and other meat products using reversed-phase-HPLC. Z Lebensm Unters Forsch 1993;197:2302. [98] Summo C, Caponio F, Paradiso VM, Pasqualone A, Gomes T. Vacuum-packed ripened sausages: evolution of oxidative and hydrolytic degradation of lipid fraction during long-term storage and inuence on the sensory properties. Meat Sci 2010;84:14751. [99] Suzzi G, Gardini F. Biogenic amines in dry fermented sausages: a review. Int J Food Microbiol 2003;88:4154. [100] Talon R, Walter D, Montel MC. Growth and effect of staphylococci and lactic acid bacteria on unsaturated free fatty acid. Meat Sci 2000;54:417. [101] Tkachenko AG, Nesterova LY, Pshenichnov K. Role of putrescine in the regulation of the expression of the oxidative stress defence genes of Escherichia coli. Microbiology 2001;70:1337. [102] Toldr F. Proteolysis and lipolysis in avour development of dry-cured meat products. Meat Sci 1998;49:S10110. [103] Toldr F, Reig M. Innovations for healthier processed meats. Trends Food Sci Technol 2011;22:51722. [104] Typpnen S, Petj E, Mattila-Sandholm T. Bioprotectives and probiotics for dry sausages review. Int J Food Microbiol 2003;83:23344. [105] Valencia I, Ansorena D, Astiasarn I. Stability of linseed oil and antioxidants containing dry fermented sausages: a study of the lipid fraction during different storage conditions. Meat Sci 2006;73:26977. [106] Vermeiren L, Devlieghere F, Debevere J. Evaluation of meat born lactic acid bacteria as protective cultures for the biopreservation of cooked meat products. Int J Food Microbiol 2004;96:14964. [107] Viljanen K. Protein oxidation and proteinlipid interactions in different food models in the presence of berry phenolics. Dissertation. EKT series 1342. Helsinki: University of Helsinki; Department of Applied Chemistry and Microbiology; 2005. [108] Vinci G, Antonelli ML. Biogenic amines: quality index of freshness in red and white meat. Food Control 2002;13:51924.

1772

D. Koozyn-Krajewska, Z.J. Dolatowski / Process Biochemistry 47 (2012) 17611772 [112] Zdolec N, Hadziosmanovic M, Kozacinski L, Cvrtila Z, Filipovic I, Skrivanko M, et al. Microbial and physicochemical succession in fermented sausages produced with bacteriocinogenic culture of Lactobacillus sakei and semi-puried bacteriocin mesenterocin Y. Meat Sci 2008;80:4807. [113] Zhang X, Kong B, Xiong L. Production of cured meat color in nitritefree Harbin red sausage by Lactobacillus fermentum fermentation. Meat Sci 2007;77:5938. [114] Zhou GH, Zhao GM. Biochemical changes during processing of traditional Jinhua ham. Meat Sci 2007;77:11420.

[109] Virgili R, Saccani G, Gabba L, Tanzi E, Soresi Bordini C. Changes of free amino acids and biogenic amines during extended ageing of Italian dry-cured ham. LWT-Food Sci Technol 2007;40:8718. A. The effect of probiotic strains on [110] Wjciak K, Dolatowski ZJ, Okon oxidative stability of cured pork meat products. Fleischwirtschaft Int 2012;27(1):1004. [111] Yeo SK, Ewe JA, Tham CSC, Liong MT. Carriers of probiotic microorganisms. In: Liong MT, editor. Probiotics, microbiology monographs 21. Berlin/Heidelberg: Springer-Verlag; 2011. p. 191220.